WE MEET AGAIN....
Hey there we meet again; if you are reading from my last blog, I am glad I have entertained you so far. So, I hope you remember my name, and if you don’t, hint – it might be in the title. More importantly, I would be glad, if you remembered my previous discussion on community service learning (CSL). To recap, I emphasized that doing CSL was more than just volunteering. It helps enhance learning in class with real life situations and builds ideas and opportunities that can allow one to essentially choose a career. I bet, you are thinking she is really trying to sell this “CSL” thing, and if you are thinking that you are actually right, I am, indeed. Inspiring or sparking the want to do something in someone, never hurts anyone. But, for now enough of why choose CSL and more of “but, why are you actually doing this again?."
What I did in the past few weeks for CSL?
As you know, I am currently working on a zebrafish courtship analysis. The analysis focuses on the behaviours of zebrafish during spawning. I am analyzing 2 hour pre-recorded videos, which were either injected with pheromones or peptides into the water tank. I looked for two courtship behaviours; quiver and tail-nose. To remind you of those behaviours, the pictures are placed to the right. These video analyses are very important as they represent data for actual research; therefore it is important to begin the real analysis with a practice video. We used our good friend excel, but it was a painful, and tedious process as we continuously paused and recorded EACH time a tail-nose/quiver occurred. Then, came BORIS, aka the life saver, but only half of our problems were solved. On another note, it may sound like a very easy task to be looking at a fish and pointing out what is a tail-nose and a quiver. But guess what, it wasn’t, in fact barely anyone who did the practice video got results close to the actual results. And, my friend this is why CSL is a great learning experience; you get exposed to problems and then you learn how to work around them. However, once we mastered the practice video, the real analysis started on the full 2 hour video. The analysis is completed at 50% of the original speed and even lower sometimes because tail-noses are extremely rapid to notice. On the other hand not many quivers occur throughout the video, however both of these behaviours aid in the final results. The video analysis takes approximately 7-8 hours per video.
TAIL-NOSE
QUIVER
We decided to change up: How did it work?
Just as of recently, there are modifications that were to the project. As explained above, the analysis is time consuming, as all times must be recorded, wherever the zebrafish shows the behaviour. Although, both of the behaviours are important in the final analysis, tail-nose is being taken out. Quivers are not as frequent, therefore a 2 hour video, may take 3-4 hours to analyze to fasten up the process. Furthermore, in quivers it is easier to tell the difference between the control experiment and the knockout experiment. Quivers are easier to recognize and are not as frequent making it easy to use them as behaviour to compare. It is hard to notice all of the tail-noses, as the data showed variability, it was easier to just drop them out of the analysis. You may be wondering, why are these behaviours so important? What are these behaviours even helpful in determining?
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The importance of the behaviours was something I touched upon last time, but I will explain it in detail now. The hypothesis tested by Kimberly Mitchell, focuses on the idea that secretoneurin (SN) is a new reproductive hormone. The experiment makes use of a knockout fish in which a targeted gene (SgII) is disrupted to determine the effect of this hormone. These behaviours will help determine if the injected SN or pheromones can rescue spawning in the knockout fish. It is often hard to see, and more importantly connect, what we do to the overall big conclusions. The data obtained from this analysis is being used to determine the impact of a new peptide on the zebrafish. The behaviours indicate that the injected pheromones indeed helped with spawning. We want these results, because it essentially shows that SN is a hormone peptide.
Future of the conducted analysis?
The main focus of the study is to analyze the effect of reproduction in vertebrates, in particular zebrafish (Danio rerio). The hormone involved in reproductive processes is luteinizing hormone (LH) which is an anterior pituitary hormone. It is responsible for gonadal development, sex hormones, and sex pheromone synthesis. It is crucial to activate this hormone in order for reproduction to occur; otherwise it leads to reduced fertility. University of Ottawa has conducted previous studies which indicate the peptide; secretoneurin (SN) stimulates the pituitary to release LH which leads to reproduction. Kimberley Mitchell is hypothesizing that SN is a new reproductive peptide hormone. The goal is to determine the role of it in reproduction in knockout fish. A knockout fish is a genetically modified fish, in which a targeted gene (SgII) is disrupted in order to determine the effect of the new reproductive hormone. SgII is a precursor protein called secretogranin, which through proteolysis (breakdown of proteins into smaller polypeptides) transforms into SN. A SgII knockout fish is used in order to eliminate the function of SgII/SN and be able to draw comparisons that conclude SN is a new reproductive hormone. The spawning videos are used to look for courtship behaviours, and will allow concluding if injected SN or pheromones can save spawning in zebrafish.
The past few weeks have been great in learning the impact of the research we are conducting. It was fun to take you on this journey with me. I hope I inspired you in some way to try to make an effort and do something out of your comfort zone. It is important to remember that a lot of layers make up an individual, one those layers may be marks, but real life experience is definitely the most important and valuable layer. Lastly, CSL has enhanced my knowledge about genetics by allowing me to connect class concepts to real life.
The main focus of the study is to analyze the effect of reproduction in vertebrates, in particular zebrafish (Danio rerio). The hormone involved in reproductive processes is luteinizing hormone (LH) which is an anterior pituitary hormone. It is responsible for gonadal development, sex hormones, and sex pheromone synthesis. It is crucial to activate this hormone in order for reproduction to occur; otherwise it leads to reduced fertility. University of Ottawa has conducted previous studies which indicate the peptide; secretoneurin (SN) stimulates the pituitary to release LH which leads to reproduction. Kimberley Mitchell is hypothesizing that SN is a new reproductive peptide hormone. The goal is to determine the role of it in reproduction in knockout fish. A knockout fish is a genetically modified fish, in which a targeted gene (SgII) is disrupted in order to determine the effect of the new reproductive hormone. SgII is a precursor protein called secretogranin, which through proteolysis (breakdown of proteins into smaller polypeptides) transforms into SN. A SgII knockout fish is used in order to eliminate the function of SgII/SN and be able to draw comparisons that conclude SN is a new reproductive hormone. The spawning videos are used to look for courtship behaviours, and will allow concluding if injected SN or pheromones can save spawning in zebrafish.